We are investigating the biosynthesis of Rous sarcoma virus (RSV) and the number and nature of virus-specific protein produced in cell-free extracts in response to added virion RNA and mRNA from transformed cells. The work encompasses the following projects to be carried out in parallel. 1. Analyze the polypeptides synthesized in wheat-germ cell-free extracts in response to 35S virion RNa, 2. Similarly, analyze polypeptides produced in response to subgenomic fragments of RSV RNA prepared from wild-type and transformation-defective virus, 3. Evaluate the effect of added virion tRNA on the above translation, 4. Determine the size and function of virus-specific RNA's synthesized in transformed cells, 5. Search for alterations in protein biosynthesis in transformed cells as compared with their normal counterparts by two-dimensional electrophoretic analysis of proteins and by examination of DNA-binding proteins. We will utilize 1. wheat-germ cell-free extracts, 2. molecular hybridization, 3. two-dimensional electrophoresis and conventional slab gel SDS-acrylamide electrophoresis of proteins, 4. conditional and non-conditional transformation mutants of RSV, 5. sucrose gradient and acrylamide gel analysis of RNA, 6. oligo (dT)-cellulose and DNA-cellulose chromatography, 7. immunoprecipitation.